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Growth Requirements of Pathogenic Leptospira

机译：致病性钩端螺旋体的生长要求

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Nutritional requirements for growth at 30 C of Leptospira pomona and L. canicola have been determined. Both pathogenic serotypes initially required bovine serum albumin (BSA) for growth in a medium (SM-4) which permitted growth of the water isolate B-16. Requirement for BSA was eliminated by (i) removing much of the apparent toxicity of free fatty acids in Tween 80 on an anion exchange column, (ii) decreasing extended lag periods observed from small inocula by incorporation of pyruvate into the medium, (iii) the addition of acetate to permit full utilization of substrate fatty acids in Tween 80, and (iv) the addition of glycerol to decrease generation times. Physiologic significance of these findings is discussed, and the possibility is suggested that apparent toxicity of fatty acids for leptospires may result from their auto-oxidation products. The resulting protein-free medium (SM-5) permitted the growth of pathogens at 30 C to high cell yields in low inocula. Highly virulent and avirulent strains from the same clone of L. canicola Moulton were used to determine additional growth requirements associated with virulence. As incubation temperatures were increased from 30 C to those of mammalian hosts, virulent cells required biotin at 35 C and higher levels of K+ and Mg2+ at 37 C. Additional Fe2+ eliminated the necessity for removing the toxicity of Tween 80 by anion exchange. Significance of these physiologic studies are discussed in relationship to virulence. The final protein-free medium (SM-6) grew highly virulent L. canicola from tissue to high yields from low inocula at 37 C with no loss in virulence over several transfers.
机译： Leptospira pomona 和 L在30 C下生长的营养需求。 canicola 已确定。两种致病血清型最初都需要牛血清白蛋白（BSA）在培养基（SM-4）中生长，该培养基允许水分离物B-16生长。通过（i）除去阴离子交换柱上吐温80中游离脂肪酸的大部分表观毒性，（ii）通过将丙酮酸盐掺入培养基中来减少从小接种物中观察到的延长的滞后时间，消除了BSA的要求。加入乙酸盐可充分利用吐温80中的底物脂肪酸，以及（iv）添加甘油以减少生成时间。讨论了这些发现的生理意义，并暗示了脂肪酸对钩藤螺的明显毒性可能是由于其自身的氧化产物所致。所得的无蛋白培养基（SM-5）使病原体在30℃下以低接种量生长至高细胞产量。来自 L的同一克隆的高毒力和无毒力菌株。 canicola Moulton用于确定与毒力相关的其他生长需求。随着孵育温度从30°C升高到哺乳动物宿主的温度，有毒细胞需要在35°C的生物素和在37°C的K + 和Mg 2 + 更高的水平。 Fe 2 + 消除了通过阴离子交换去除Tween 80毒性的必要性。这些生理学研究的意义与毒力的关系进行了讨论。最终的无蛋白培养基（SM-6）增长了高毒性的 L。在低至37℃的接种量下从组织到高产量的Canicola ，在几次转移中均无毒力损失。

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